Minims eye drops

Adult Cx3cr1 þ/eGFP mice (n ¼ 27 in total) were anaesthetized using a mixture of ketamine (67 mg/kg) and xylazine (13 mg/ kg) and the ocular surface further anaesthetized with topical proxymetacaine (Alcaine, 0.5%; Alcon Laboratories, Frenchs Forest, NSW, Australia). Following dilation of the pupil with 0.5% tropicamide (Mydriacyl; Alcon Laboratories) and 10% phenylephrine hydrochloride (Minims Eye Drops; Bausch & Lomb, Macquarie Park, NSW, Australia), single intravitreal injections were made 1 mm posterior to the limbus using a 31-G needle attached to a 10-lL Hamilton syringe. Mice received 1-lL injections of 10 mM AngII (Sigma-Aldrich Corp., final concentration at retina given a 5-lL vitreal volume of 2 mM 30 ), with the fellow eye receiving a 1-lL injection of vehicle control (phosphate-buffered saline [PBS]). Directly after intravitreal injection, fluorescein angiography was conducted on a subset of mice (n ¼ 3) to assess changes in blood vessel caliber in response to AngII. Mice received a 100-lL intraperitoneal injection of 0.2% sodium fluorescein (Alcon Laboratories), and the retinal fundi imaged with a Micron III retinal imaging system (Phoenix Research Laboratories, Pleasanton, CA, USA) for 20 minutes as previously described. 31 Fundus images were collected and processed using the Micron III software (Stream-Pix 5.0; NorPix, Inc., Quebec, Canada).